Status
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Gene information
General information
TC033450
Former TC numbers

If one gene is splitted or genes are merged, the original TC numbers are listed. One listed former TC number indicates the previous gene has be splitted, serveral former TC number represents a merged gene.

TC015795,TC015796
Gene

The sequence from the iB fragment is matched against the genes from the official gene set. The matche(s) is/are visualizied in a genome browser.

NC_007421.3: 8297911..8422372
Sequence information
Transcripts / Proteins (2) Get mRNA sequence
Get CDS sequence
Get protein sequence
Ortholog information (from OrthoDB)

The information about the homolog genes in Drosophila is provided by OrthoDB http://www.orthodb.org/. The homologs are sorted according to their similarity score. For an orthology statement additional analysis are required.
The data basis for the orthologs is OrthoDB v9 containing OGS3 for Tribolium castaneum and 2015_04 (r6.07) for Drosophila melanogaster.

Closest fly homologs
    TC033450
    iBeetle screen iB_05895

    Attention! iBeetle is a first pass screen. Hence, the documented phenotypes need to be confirmed by independent experiments using non-overlapping dsRNA fragments! Phenotypes with a penetrance > 50% are frequently reproduced. See Schmitt-Engel et al. 2015 for details on reproducibility.

    iB sequence
    Phenotype after pupal injection
    Usually 10 injected animals

    Pupal injection procedure

    Day 0: 10 female pupae of the pBA19 strain (muscle enhancer trap line) were injected with dsRNA.

    3 days post injection (dpi): Hatch control: Pupal and adult lethality as well as metamorphosis defects (molting, eclosion) were documented. For mating, 4 males of the black strain were added.

    9 dpi: First egg-lay was collected and incubated for cuticle analysis. Adult lethality and egg production (reduced/ no egg-lay) was documented.

    11 dpi: Second egg-lay was collected and incubated for embryonic muscle analysis. Adult lethality and egg production (reduced/ no egg-lay) were documented.

    Note: The adult morphology was not analyzed systematically at 3/9/11 dpi. Only obvious phenotypes, visible without magnification, have been annotated.

    13 dpi: Egg productivity and Ovary analysis
    The percentage of hatched larvae was documented and not hatched larvae/ eggs were embedded for cuticle analysis (15 dpi). In case of a reduction of egg production, 4 injected females were dissected to analyze the gross morphology of the ovaries.

    14 dpi: Analysis of embryonic musculature and early embryonic development
    Offspring of the injected females (hatched and not hatched larvae/ eggs) were analysed for embryonic lethality and muscle defects.

    15 dpi: Analysis of larval instar 1 cuticle
    Offspring of injected females were analysed and cuticle phenotypes were annotated.

    22 dpi: Stink gland analysis
    Documentation of defects in abdominal and thoracic stink glands (colour, size, content) of the injected femals.

    Terms used in the pupal injection procedure

    larval pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    empty eggs
    Just empty shells are visible, no cuticle has been developed.
    strong defects
    L1 cuticle larvae which show severe defects. These strong defects are subdivided into three categories
    strongly fragmented (cuticle crumbs)
    No segments or appendages are identifiable, only residues of cuticle and bristles can be identified.
    cuticle remnants (with unidentifiable segments)
    Segments or parts of appendages are recognizable, but identity is not clear.
    cuticle remnants (with some identifiable segments)
    At least one segment or appendage is clearly regonizable.
    number of eggs on slide
    affected embryos usually (not always) do not hatch. Only the not hatched were used for cuticle analysis.
    For the embryonic muscle analysis the hatched as well as the not hatched were analysed.
    total number of affected eggs/embryos/larvae
    Summary of all different phenotypes/ phenotypic series which were annotated.
    inside-out
    The L1 larva shows a partial or complete inversion., e.g. bristles, appendages or parts of the abdominal segments are inverted into the interior of the cuticle.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    Please see the help page for more information
    Metamorphosis and survival

    Lethality 11 days after pupal injection: 50.0% (includes death as pupa, adult)

    eclosion not fulfilled (with pupal and adult features) - Number of animals showing the phenotype: 2 (3 dpi)
    Days post injection


    adult lethality - Number of animals showing the phenotype: 3 (9 dpi)
    Days post injection


    Analysis of embryonic musculature and early development
    number of eggs/embryos/larvae on slide: > 50 larval musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 30% - 50%

    leg musculature pattern irregular - Penetrance (percentage of animals/eggs on slide): 50% - 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): 50% - 80%
    remarks: muscle pattern potentially obscured by segmentation defect,
    Analysis of larval stage 1 cuticle
    number of eggs/embryos/larvae on slide: > 50
    maxilla shape irregular - Penetrance (percentage of animals/eggs on slide): more than 80%

    legs shortened - Penetrance (percentage of animals/eggs on slide): more than 80%

    legs femur potentially fused - Penetrance (percentage of animals/eggs on slide): more than 80%

    legs tibiotarsus potentially fused - Penetrance (percentage of animals/eggs on slide): more than 80%
    urogomphi shortened - Penetrance (percentage of animals/eggs on slide): more than 80%

    affected eggs/embryos/larvae - Penetrance (percentage of animals/eggs on slide): more than 80%
    Phenotype after larval injection
    Usually 10 injected animals

    Larval injection procedure

    Day 0: 10 female L5-6 larvae of the D17Xhom strain (females express RFP in the eyes and in the CNS; pupae express GFP in the thorax) were injected with dsRNA.

    11 days post injection (dpi): Pupal morphology Documentation of larval/ pupal death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of pupal head and appendages, pupal thorax, pupal thoracic musculature, pupal legs, pupal wings, pupal abdomen and pupal genital lobes.

    16 dpi: Adult morphology Documentation of larval/ pupal and adult death and defects of metamorphosis (eclosion, delayed development). Screening for alterations of adult head and appendages, adult thorax, adult elytra and hindwings, adult legs and cuticle properties. For mating 4 males of the black strain were added.

    19 dpi: Sieving and egg-lay Documentation of adult lethality and enduring metamorphosis defects.

    22 dpi: Ovary analysis (Fertility) In case of a reduction of egg production 4 females were dissected and the gross morphology of the ovaries were analysed.

    41 dpi: Fertility and stink gland analysis Documentation of defects in abdominal and thoracic stink glands development (colour, size, content) of the injected females. Check for fertility of injected larvae (no or decreased offspring).

    Terms used in the larval injection procedure

    pupal/ adult pantagmatic defects
    At least two tagmata (head, thorax, abdomen or terminus) show similar/ comparable defects.
    pupal molt not fulfilled
    The hatching from the larva to the pupa is interrupted. Either it dies as prepupa or the phenotype shows larval as well as pupal features.
    eclosion not fulfilled
    The emergence of the adult from the pupa stage is interrupted. This phenotype shows pupal as well as adult features.
    starved
    Dissected females show a strong resorption of fatbody predominately in the abdomen and the oogenesis is blocked. These are considered as eventually lethal phenotypes and the blocked oogenesis probably a secondary defect due to starvation.
    Metamorphosis and survival

    Lethalities 11 days after larval injection: 40.0% (includes death as larva, prepupa, pupa)
    Lethalities 22 days after larval injection: -1.0% (includes death as larva, prepupa, pupa, adult)

    RNAi sequence
    Primers
    Left: Right:
    Protein sequence
    • >TC033450 RA
      MRFDTTLEP PLDAKVKTT MAYHPFLQL PRQTDFSVS 
      SLLTAASSS NPTSPPAPG FFPSAALAA FGSQASHGC 
      YPGTLIPKI SHPHPHHAL SGHPYTTAE DVVLAAVAA 
      HNHHPGMVR PLRAIQPEE DGVVDDPKV TLEGKDLWE 
      KFHKLGTEM VITKSGRQM FPQMKFRVS GLDLKAKYI 
      LLLDIVAAD DYRYKFHNS RWMVAGKAD PEMPKRMYI 
      HPDSPSTGE QWMQKVVSF HKLKLTNNI SDKHGFLKE 
      KAVDFLTIL NSMHKYQPR FHLVRANDI LKLPYSTFR 
      TYVFKETEF IAVTAYQNE KITQLKIDN NPFAKGFRD 
      TGAGKREKK QAMLAQRHT DDKTSHPLP ARMSADSST 
      VSKTDDSEL DVVGTADSP RPSLHASTP IASSLNHSG 
      PEDDSLRRR LHDDSGSES SCSESPGAF RPRPSSPKE 
      TTAGPSNPS GEYPSPNIS VGPPIHPPP HLLPYLYPP 
      GLYPGAGGP PFAPHLNLF SGAHAPGMN PSLLFNAQL 
      ALAAQHPAL FSHYQNLAH PNPHHLSPT SPLHNHLKG 
      GHRFTPYTL PTSNSGVSS SPLGSAFET VTPSSHTRS 
      LSNSPSGKL RGGSMSPPV RPSTASPKP EAATSPSDL 
      KSIEKMVNG LDVKQNEDN DK
    • >TC033450 RB
      MRFDTTLEP PLDAKVKTT MAYHPFLQL PRQTDFSVS 
      SLLTAASSS NPTSPPAPG FFPSAALAA FGSQASHGC 
      YPGTLIPKI SHPHPHHAL SGHPYTTAE DVVLAAVAA 
      HNHHPGMVR PLRAIQPEE DGVVDDPKV TLEGKDLWE 
      KFHKLGTEM VITKSGRQM FPQMKFRVS GLDLKAKYI 
      LLLDIVAAD DYRYKFHNS RWMVAGKAD PEMPKRMYI 
      HPDSPSTGE QWMQKVVSF HKLKLTNNI SDKHGFTIL 
      NSMHKYQPR FHLVRANDI LKLPYSTFR TYVFKETEF 
      IAVTAYQNE KITQLKIDN NPFAKGFRD TGAGKREKK 
      QAMLAQRHT DDKTSHPLP ARMSADSST VSKTDDSEL 
      DVVGTADSP RPSLHASTP IASSLNHSG PEDDSLRRR 
      LHDDSGSES SCSESPGAF RPRPSSPKE TTAGPSNPS 
      GEYPSPNIS VGPPIHPPP HLLPYLYPP GLYPGAGGP 
      PFAPHLNLF SGAHAPGMN PSLLFNAQL ALAAQHPAL 
      FSHYQNLAH PNPHHLSPT SPLHNHLKG GHRFTPYTL 
      PTSNSGVSS SPLGSAFET VTPSSHTRS LSNSPSGKL 
      RGGSMSPPV RPSTASPKP EAATSPSDL KSIEKMVNG 
      LDVKQNEDN DK
    CDS sequence
    • >TC033450 RA
      atgcgtttc gatacgacc ctggagcca cccctcgac 
      gccaaagtg aagaccacg atggcctac caccccttc 
      ctgcagctg ccgcgccag acggacttc agcgtcagc 
      tcgctgctc accgccgcc tccagcagc aaccccacg 
      tcgccgccc gcccccggc ttcttcccg tcggcggcg 
      ctcgccgcc ttcggctcg caggcctcg cacggctgc 
      tacccgggc acgctcata cccaagatc agccatccg 
      cacccgcat cacgccctc tcgggacac ccctacacc 
      accgccgag gacgtggtg ctggccgcc gtcgccgcg 
      cacaaccac cacccgggg atggtcagg ccgctgagg 
      gcgatccag ccggaggag gatggggtg gtcgacgat 
      cccaaagtc acacttgaa gggaaagat ctctgggaa 
      aagtttcat aaactcggc acggaaatg gtcatcacc 
      aaaagtggg agacaaatg tttccccag atgaagttc 
      agagtgtcc ggtttggac ctgaaagcc aaatacata 
      ctcctgttg gacatagtc gccgctgac gactacaga 
      tacaaattc cataacagc cggtggatg gtggcgggc 
      aaggccgac cccgaaatg cccaaacgg atgtacatc 
      cacccagac agcccctcg accggcgag cagtggatg 
      cagaaagtc gtctccttc cacaagctc aagctcacc 
      aacaacatc tcggacaag cacggcttt ttgaaagaa 
      aaagcggtt gatttcctg acgatactc aactccatg 
      cacaaatac cagcctcgg tttcacctc gttagggcc 
      aacgacatt ttgaagttg ccctactcg acgtttcgg 
      acgtacgtc ttcaaggaa acggagttc atcgcagtg 
      acagcgtac caaaacgaa aagataacg cagctcaag 
      atcgacaac aatccgttc gcgaagggg ttcagggat 
      acgggggcg ggcaagagg gagaaaaag caggcgatg 
      ctggcgcag agacacacc gatgataag acgtcacat 
      ccgcttcct gcgaggatg tcggcggac agcagcacc 
      gtatcaaag actgatgac tcggaattg gatgttgtg 
      gggacggcg gacagtccg aggccgtcc ctgcacgcc 
      tccacgccg attgcgtct tcgcttaat cattccggc 
      cccgaagac gactccctt cgccggcgc ctccacgac 
      gactccggc tctgaatcc tcgtgttca gaatccccc 
      ggagccttc cggccgcgg ccctcctcc cccaaggag 
      accaccgcc ggcccctcc aacccctct ggcgagtac 
      ccctcgccc aacatcagc gtgggcccc ccaatccac 
      cccccgcca cacctgctc ccgtacctg tacccccct 
      ggcctgtac cccggcgcg gggggcccg cctttcgcc 
      ccccatttg aacctcttt tccggggcg cacgccccc 
      ggcatgaac cccagcctc ctcttcaac gctcaacta 
      gccctcgcc gcccaacac cccgccctc ttctcccac 
      taccaaaac ctagcgcac cccaacccc caccacttg 
      tcccccacc tccccgctg cataaccac ctcaagggg 
      gggcacagg ttcaccccc tacacgctg cccacctcc 
      aactcgggg gtgagttcg tcccccttg gggagtgcg 
      ttcgagacg gtcaccccc agctcgcac acgaggagt 
      ttgagtaat tctccctcg gggaagctg aggggcggg 
      tcgatgtcg cccccagtg aggcccagt accgcctcc 
      cccaagccg gaggcggcg acctccccc agcgacctc 
      aagagtata gaaaaaatg gtaaatgga ctcgatgtt 
      aagcaaaac gaagacaat gacaagtag
    • >TC033450 RB
      atgcgtttc gatacgacc ctggagcca cccctcgac 
      gccaaagtg aagaccacg atggcctac caccccttc 
      ctgcagctg ccgcgccag acggacttc agcgtcagc 
      tcgctgctc accgccgcc tccagcagc aaccccacg 
      tcgccgccc gcccccggc ttcttcccg tcggcggcg 
      ctcgccgcc ttcggctcg caggcctcg cacggctgc 
      tacccgggc acgctcata cccaagatc agccatccg 
      cacccgcat cacgccctc tcgggacac ccctacacc 
      accgccgag gacgtggtg ctggccgcc gtcgccgcg 
      cacaaccac cacccgggg atggtcagg ccgctgagg 
      gcgatccag ccggaggag gatggggtg gtcgacgat 
      cccaaagtc acacttgaa gggaaagat ctctgggaa 
      aagtttcat aaactcggc acggaaatg gtcatcacc 
      aaaagtggg agacaaatg tttccccag atgaagttc 
      agagtgtcc ggtttggac ctgaaagcc aaatacata 
      ctcctgttg gacatagtc gccgctgac gactacaga 
      tacaaattc cataacagc cggtggatg gtggcgggc 
      aaggccgac cccgaaatg cccaaacgg atgtacatc 
      cacccagac agcccctcg accggcgag cagtggatg 
      cagaaagtc gtctccttc cacaagctc aagctcacc 
      aacaacatc tcggacaag cacggcttt acgatactc 
      aactccatg cacaaatac cagcctcgg tttcacctc 
      gttagggcc aacgacatt ttgaagttg ccctactcg 
      acgtttcgg acgtacgtc ttcaaggaa acggagttc 
      atcgcagtg acagcgtac caaaacgaa aagataacg 
      cagctcaag atcgacaac aatccgttc gcgaagggg 
      ttcagggat acgggggcg ggcaagagg gagaaaaag 
      caggcgatg ctggcgcag agacacacc gatgataag 
      acgtcacat ccgcttcct gcgaggatg tcggcggac 
      agcagcacc gtatcaaag actgatgac tcggaattg 
      gatgttgtg gggacggcg gacagtccg aggccgtcc 
      ctgcacgcc tccacgccg attgcgtct tcgcttaat 
      cattccggc cccgaagac gactccctt cgccggcgc 
      ctccacgac gactccggc tctgaatcc tcgtgttca 
      gaatccccc ggagccttc cggccgcgg ccctcctcc 
      cccaaggag accaccgcc ggcccctcc aacccctct 
      ggcgagtac ccctcgccc aacatcagc gtgggcccc 
      ccaatccac cccccgcca cacctgctc ccgtacctg 
      tacccccct ggcctgtac cccggcgcg gggggcccg 
      cctttcgcc ccccatttg aacctcttt tccggggcg 
      cacgccccc ggcatgaac cccagcctc ctcttcaac 
      gctcaacta gccctcgcc gcccaacac cccgccctc 
      ttctcccac taccaaaac ctagcgcac cccaacccc 
      caccacttg tcccccacc tccccgctg cataaccac 
      ctcaagggg gggcacagg ttcaccccc tacacgctg 
      cccacctcc aactcgggg gtgagttcg tcccccttg 
      gggagtgcg ttcgagacg gtcaccccc agctcgcac 
      acgaggagt ttgagtaat tctccctcg gggaagctg 
      aggggcggg tcgatgtcg cccccagtg aggcccagt 
      accgcctcc cccaagccg gaggcggcg acctccccc 
      agcgacctc aagagtata gaaaaaatg gtaaatgga 
      ctcgatgtt aagcaaaac gaagacaat gacaagtag
    mRNA sequence
    • >TC033450 RA
      cacgtgacg tgtccgcta atgaaaacc gtgtgtttt 
      tgcaaatgt gtatgtgtg tgtgtgtgc ggcgcttaa 
      aaaggtcga gcgttggtg gcgtctgaa cagcatgcg 
      gtgaattca tgcatatgc gtcccgttt ggtaaagga 
      gcattcagt cctggcaac atcaatgtc gcctcagca 
      gaaaaagga aaccgtgtg acaaaaagc actcgtaac 
      gcctccata ggcataacc tcctaactc ctgcccacc 
      gtaagtgtg aaaagtggg ggcgaaatg tcgccatcg 
      gattggcgg agccgcttg agctgacac cctcgcgac 
      caatcccat cgcgtccgt gtcattaac acagtcagt 
      gtcgaatct gcgggtttc ctgctatag tcgtcgctc 
      acacttaac ccgggaagg aggtgtcag acttcacac 
      tcaagccac cccgcgttc aggacctac tcagtgcag 
      tgcgtggtg cagtgcggt gatgtgata tacagggcc 
      ggcgtcgga ccatgcgtt tcgatacga ccctggagc 
      cacccctcg acgccaaag tgaagacca cgatggcct 
      accacccct tcctgcagc tgccgcgcc agacggact 
      tcagcgtca gctcgctgc tcaccgccg cctccagca 
      gcaacccca cgtcgccgc ccgcccccg gcttcttcc 
      cgtcggcgg cgctcgccg ccttcggct cgcaggcct 
      cgcacggct gctacccgg gcacgctca tacccaaga 
      tcagccatc cgcacccgc atcacgccc tctcgggac 
      acccctaca ccaccgccg aggacgtgg tgctggccg 
      ccgtcgccg cgcacaacc accacccgg ggatggtca 
      ggccgctga gggcgatcc agccggagg aggatgggg 
      tggtcgacg atcccaaag tcacacttg aagggaaag 
      atctctggg aaaagtttc ataaactcg gcacggaaa 
      tggtcatca ccaaaagtg ggagacaaa tgtttcccc 
      agatgaagt tcagagtgt ccggtttgg acctgaaag 
      ccaaataca tactcctgt tggacatag tcgccgctg 
      acgactaca gatacaaat tccataaca gccggtgga 
      tggtggcgg gcaaggccg accccgaaa tgcccaaac 
      ggatgtaca tccacccag acagcccct cgaccggcg 
      agcagtgga tgcagaaag tcgtctcct tccacaagc 
      tcaagctca ccaacaaca tctcggaca agcacggct 
      ttttgaaag aaaaagcgg ttgatttcc tgacgatac 
      tcaactcca tgcacaaat accagcctc ggtttcacc 
      tcgttaggg ccaacgaca ttttgaagt tgccctact 
      cgacgtttc ggacgtacg tcttcaagg aaacggagt 
      tcatcgcag tgacagcgt accaaaacg aaaagataa 
      cgcagctca agatcgaca acaatccgt tcgcgaagg 
      ggttcaggg atacggggg cgggcaaga gggagaaaa 
      agcaggcga tgctggcgc agagacaca ccgatgata 
      agacgtcac atccgcttc ctgcgagga tgtcggcgg 
      acagcagca ccgtatcaa agactgatg actcggaat 
      tggatgttg tggggacgg cggacagtc cgaggccgt 
      ccctgcacg cctccacgc cgattgcgt cttcgctta 
      atcattccg gccccgaag acgactccc ttcgccggc 
      gcctccacg acgactccg gctctgaat cctcgtgtt 
      cagaatccc ccggagcct tccggccgc ggccctcct 
      cccccaagg agaccaccg ccggcccct ccaacccct 
      ctggcgagt acccctcgc ccaacatca gcgtgggcc 
      ccccaatcc accccccgc cacacctgc tcccgtacc 
      tgtaccccc ctggcctgt accccggcg cggggggcc 
      cgcctttcg ccccccatt tgaacctct tttccgggg 
      cgcacgccc ccggcatga accccagcc tcctcttca 
      acgctcaac tagccctcg ccgcccaac accccgccc 
      tcttctccc actaccaaa acctagcgc accccaacc 
      cccaccact tgtccccca cctccccgc tgcataacc 
      acctcaagg gggggcaca ggttcaccc cctacacgc 
      tgcccacct ccaactcgg gggtgagtt cgtccccct 
      tggggagtg cgttcgaga cggtcaccc ccagctcgc 
      acacgagga gtttgagta attctccct cggggaagc 
      tgaggggcg ggtcgatgt cgcccccag tgaggccca 
      gtaccgcct cccccaagc cggaggcgg cgacctccc 
      ccagcgacc tcaagagta tagaaaaaa tggtaaatg 
      gactcgatg ttaagcaaa acgaagaca atgacaagt 
      agtgaaagt gatcctaag ttttagttg gccgtactg 
      atgatatat acggggtgt ttttgcaat agattcttt 
      tggattttt gtgattttt gcacatgaa attaccgta 
      aagtgcttt tcttattgt agcttacaa aaaaagtac 
      aaaatgtat gtgaattga ggcagattt tttcgttta 
      atccgacct tatttggtg ctatgctca ttctgtttg 
      accctagaa aggaagaaa gtctggaag aggggtcac 
      ttttagact ttgttcctc ctcaattag tgatcctgt 
      agactgttg aagcttttg tataataat aatgatatt 
      tactaagag acacgatag aagagttaa tgttccttt 
      tactattat tgcaattaa tttaaataa ttatgtaaa 
      tagtgtttg tacgtaaac gcgtaaaca tctgtatat 
      aatggttgt tttttgtgt tttatgaag aaaaactgt 
      aaattgtaa attgctcct ataactact agaactgta 
      tattagaac tcaaaatta atttatatt tctaccaaa 
      gaatttcga tggtttttg caccaacag ctgggcaaa 
      aaccaccag aaatagtga atatagcaa taaagaagt 
      acataaacc ataaacctt ctctcgaaa ctctcaaca 
      ttgtacata aatatagtt taaatgtac atttcttta 
      ttgacatgc ttttcaaat cctacttct tctgtttat 
      atcgttgag cagtaccag cgttaaaat aacttgttt 
      ctatttatt atcaaaagt tttatattc aaatactat 
      catcgctta acgctggta cttggaaat tattaaaat 
      ctgtattaa agtagaaga tgtgaaatt tgtcttgtg 
      tatatgtgg aatactttt aagaattaa acgatattc 
      atacat
    • >TC033450 RB
      cacgtgacg tgtccgcta atgaaaacc gtgtgtttt 
      tgcaaatgt gtatgtgtg tgtgtgtgc ggcgcttaa 
      aaaggtcga gcgttggtg gcgtctgaa cagcatgcg 
      gtgaattca tgcatatgc gtcccgttt ggtaaagga 
      gcattcagt cctggcaac atcaatgtc gcctcagca 
      gaaaaagga aaccgtgtg acaaaaagc actcgtaac 
      gcctccata ggcataacc tcctaactc ctgcccacc 
      gtaagtgtg aaaagtggg ggcgaaatg tcgccatcg 
      gattggcgg agccgcttg agctgacac cctcgcgac 
      caatcccat cgcgtccgt gtcattaac acagtcagt 
      gtcgaatct gcgggtttc ctgctatag tcgtcgctc 
      acacttaac ccgggaagg aggtgtcag acttcacac 
      tcaagccac cccgcgttc aggacctac tcagtgcag 
      tgcgtggtg cagtgcggt gatgtgata tacagggcc 
      ggcgtcgga ccatgcgtt tcgatacga ccctggagc 
      cacccctcg acgccaaag tgaagacca cgatggcct 
      accacccct tcctgcagc tgccgcgcc agacggact 
      tcagcgtca gctcgctgc tcaccgccg cctccagca 
      gcaacccca cgtcgccgc ccgcccccg gcttcttcc 
      cgtcggcgg cgctcgccg ccttcggct cgcaggcct 
      cgcacggct gctacccgg gcacgctca tacccaaga 
      tcagccatc cgcacccgc atcacgccc tctcgggac 
      acccctaca ccaccgccg aggacgtgg tgctggccg 
      ccgtcgccg cgcacaacc accacccgg ggatggtca 
      ggccgctga gggcgatcc agccggagg aggatgggg 
      tggtcgacg atcccaaag tcacacttg aagggaaag 
      atctctggg aaaagtttc ataaactcg gcacggaaa 
      tggtcatca ccaaaagtg ggagacaaa tgtttcccc 
      agatgaagt tcagagtgt ccggtttgg acctgaaag 
      ccaaataca tactcctgt tggacatag tcgccgctg 
      acgactaca gatacaaat tccataaca gccggtgga 
      tggtggcgg gcaaggccg accccgaaa tgcccaaac 
      ggatgtaca tccacccag acagcccct cgaccggcg 
      agcagtgga tgcagaaag tcgtctcct tccacaagc 
      tcaagctca ccaacaaca tctcggaca agcacggct 
      ttacgatac tcaactcca tgcacaaat accagcctc 
      ggtttcacc tcgttaggg ccaacgaca ttttgaagt 
      tgccctact cgacgtttc ggacgtacg tcttcaagg 
      aaacggagt tcatcgcag tgacagcgt accaaaacg 
      aaaagataa cgcagctca agatcgaca acaatccgt 
      tcgcgaagg ggttcaggg atacggggg cgggcaaga 
      gggagaaaa agcaggcga tgctggcgc agagacaca 
      ccgatgata agacgtcac atccgcttc ctgcgagga 
      tgtcggcgg acagcagca ccgtatcaa agactgatg 
      actcggaat tggatgttg tggggacgg cggacagtc 
      cgaggccgt ccctgcacg cctccacgc cgattgcgt 
      cttcgctta atcattccg gccccgaag acgactccc 
      ttcgccggc gcctccacg acgactccg gctctgaat 
      cctcgtgtt cagaatccc ccggagcct tccggccgc 
      ggccctcct cccccaagg agaccaccg ccggcccct 
      ccaacccct ctggcgagt acccctcgc ccaacatca 
      gcgtgggcc ccccaatcc accccccgc cacacctgc 
      tcccgtacc tgtaccccc ctggcctgt accccggcg 
      cggggggcc cgcctttcg ccccccatt tgaacctct 
      tttccgggg cgcacgccc ccggcatga accccagcc 
      tcctcttca acgctcaac tagccctcg ccgcccaac 
      accccgccc tcttctccc actaccaaa acctagcgc 
      accccaacc cccaccact tgtccccca cctccccgc 
      tgcataacc acctcaagg gggggcaca ggttcaccc 
      cctacacgc tgcccacct ccaactcgg gggtgagtt 
      cgtccccct tggggagtg cgttcgaga cggtcaccc 
      ccagctcgc acacgagga gtttgagta attctccct 
      cggggaagc tgaggggcg ggtcgatgt cgcccccag 
      tgaggccca gtaccgcct cccccaagc cggaggcgg 
      cgacctccc ccagcgacc tcaagagta tagaaaaaa 
      tggtaaatg gactcgatg ttaagcaaa acgaagaca 
      atgacaagt agtgaaagt gatcctaag ttttagttg 
      gccgtactg atgatatat acggggtgt ttttgcaat 
      agattcttt tggattttt gtgattttt gcacatgaa 
      attaccgta aagtgcttt tcttattgt agcttacaa 
      aaaaagtac aaaatgtat gtgaattga ggcagattt 
      tttcgttta atccgacct tatttggtg ctatgctca 
      ttctgtttg accctagaa aggaagaaa gtctggaag 
      aggggtcac ttttagact ttgttcctc ctcaattag 
      tgatcctgt agactgttg aagcttttg tataataat 
      aatgatatt tactaagag acacgatag aagagttaa 
      tgttccttt tactattat tgcaattaa tttaaataa 
      ttatgtaaa tagtgtttg tacgtaaac gcgtaaaca 
      tctgtatat aatggttgt tttttgtgt tttatgaag 
      aaaaactgt aaattgtaa attgctcct ataactact 
      agaactgta tattagaac tcaaaatta atttatatt 
      tctaccaaa gaatttcga tggtttttg caccaacag 
      ctgggcaaa aaccaccag aaatagtga atatagcaa 
      taaagaagt acataaacc ataaacctt ctctcgaaa 
      ctctcaaca ttgtacata aatatagtt taaatgtac 
      atttcttta ttgacatgc ttttcaaat cctacttct 
      tctgtttat atcgttgag cagtaccag cgttaaaat 
      aacttgttt ctatttatt atcaaaagt tttatattc 
      aaatactat catcgctta acgctggta cttggaaat 
      tattaaaat ctgtattaa agtagaaga tgtgaaatt 
      tgtcttgtg tatatgtgg aatactttt aagaattaa 
      acgatattc atacat
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